Bioactive compositons having skin anti aging activity

ABSTRACT

The present invention relates to bioactive compositions comprising blends of camellia and feverfew serum fractions and/or kelp and parsley serum fractions.

FIELD OF THE INVENTION

The present invention relates to the field of topical applications ofskin care compositions to the skin. The invention further relates toskin care compositions comprising blends of camellia and feverfew serumfractions and/or kelp and parsley serum fractions. The invention alsorelates to methods for increasing melanin production in skin bytopically applying the skin care composition to areas thereof in orderto stimulate one or more steps in melanin synthesis.

BACKGROUND OF THE INVENTION

Human skin comprises three principal layers: the epidermis, the dermis,and the subcutaneous fat layer. The epidermis comprises four layers(from external to internal): the stratum corneum, the granular layer,the spiny layer, and the basal layer. A separate fifth layer, thestratum lucidum, may be present between the stratum corneum and granularlayer. The basal layer produces cells which gradually migrate outward toform the other epidermal layers. As these cells migrate outward, theylose their central nucleus and start to produce skin proteins (keratins)and fats (lipids). These cells are identified as keratinocytes whenpresent in the upper layers of the epidermis. Melanocytes are anotherclass of cells located in the basal layer of the epidermis. Melanocytesare responsible for the production of melanin, which is primary factorin skin pigmentation.

Melanin is produced by a complex set of reactions within the melanocyteinvolving, at a basic level, the enzyme tyrosinase and L-tyrosine as asubstrate. Tyrosinase catalyzes the conversion of L-tyrosine to DOPA(L-3,4-dihydroxyphenylalanine) and of DOPA to dopaquinone. Dopaquinoneundergoes further conversion to form melanin. Melanin aggregates inorganelles known as the melanosomes which are transferred tokeratinocytes along slender filaments of the melanocyte known asdendrites. There are approximately 1500 gene products expressed inmelanosomes with 600 of them being expressed at any given time and 100of them believed to be unique to the melanosome. In addition, there aremany regulatory elements involved in signaling, in the transport ofmelanosomes within the melanocyte, and in the transfer of melanosomes tothe keratinocytes.

The production of melanin can be triggered by a variety of external andinternal events. For example, melanocytes produce additional melaninwhen skin is subjected to UV radiation. The melanin is then transportedvia melanasomes to the keratinocytes, which then leaves the skin with apigmented or “tanned” appearance. However, chronic UV exposure may leadto more subtle changes in skin tone. Often these changes are describedas uneven tone or as a mottled appearance. Moreover,inflammation-related skin disorders such as atopic dermatitis orpost-inflammatory hypopigmentation can also leave the skin with uneventone appearance.

On the other hand, melanin is able to absorb electromagnetic irradiationespecially the wavelengths of UV radiation. Furthermore, melanin acts asa scavenger for radicals. Thus, melanin is generally considered as anatural skin filter for UV radiation. Thus, there is a desire to providecompositions and methods of treatment that can improve the appearance ofunevenly toned skin and provide a uniformly tanned appearance, which mayprovide an aesthetic benefit (e.g., perceived younger appearance thanchronological age) in addition to providing protection against UVradiation-induced skin damage.

Over recent years, consumers have increasingly demanded “natural”cosmetic products. As a result, cosmetic manufacturers have incorporatedmore plant-based materials into their cosmetic formulations. Althoughvarious plants have been used for hundreds or even thousands of yearsfor a variety of reputed indications, until recent times it has not beenpossible to clinically verify purported effectiveness or to identify newpotential uses based upon the underlying science of the plant'sbioactivity. With recent advances in science, researchers are now betterable to assess the efficacy and/or potential new uses for plants thatuntil recently were only supported by folklore. Because of the newnessof the science, and because the number of plants that could potentiallybe utilized as cosmetic bioactives is so immense, the vast majority ofplants have not yet been fully investigated.

Many of the methods used for extracting botanical components from plantsinvolve techniques that are harmful to the plant tissue compositionand/or the bioactive components of interest contained in that tissue.Consequently, traditional extraction methods often fail to deliver thefull spectrum of activities that exist within a plant cell and thus thefull potential of botanical-based cosmetic formulations is not realized.In addition, many traditional extraction methods utilize harsh chemicalsolvents, which are not “natural” and thus are materials that consumerswant to avoid applying to their skin. Furthermore, these solvent-basedprocesses produce toxic chemical wastes that can harm the environment ifnot properly handled and disposed of as hazardous waste.

Moreover, just because a material is “natural” does not guarantee thatit is free from undesired substances that would make the materialsuitable for use on skin, however. For example, many plants containphotosensitizers such as pheophorbides and/or contact allergens such asproteins. At levels naturally found in many common plants, pheophorbidesand/or proteins do not cause concern for most people. However, whenplant materials are condensed to a highly concentrated form, such asthrough extraction, these materials can be present at levels that causeskin irritation and allergic reactions, including rashes. Even whenthese materials are present at their natural levels, however, there arestill many sensitive individuals who experience negative skin reactions.

Furthermore, as demands for natural products have increased, so haveconcerns about protecting earth's natural resources. Many of the“natural” ingredients that consumers desire are derived frombioresources that are depleted and/or destroyed when harvested for usein consumer products. Thus, consumers' desire for natural, moreearth-friendly products can ironically lead to the destruction of thevery bioresources they aim to preserve. Thus, there is a need fornatural bioactive botanical compositions that maintain their spectrum ofdesired bioactivity, are suitable for topical skin application, and arenot prepared using harsh chemical solvents. Furthermore, there is a needfor cosmetic compositions containing such bioactives that are effectivefor improving the appearance of skin pigmentation. In addition, there isa need for such bioactive materials that can be harvested and processedin an ecologically sound, sustainable manner.

These and other objects of the present invention will become apparent inlight of the following disclosure.

SUMMARY OF THE INVENTION

The present invention relates to skin care compositions and methods thatcan help increase melanin production in skin by topically applying theskin care composition to areas thereof in order to stimulate one or moresteps in melanin synthesis, which may provide an aesthetic benefit(e.g., perceived younger appearance than chronological age) in additionto providing protection against UV radiation-induced skin damage.According to an embodiment, a skin care composition suitable fordarkening regions of mammalian skin is provided. The skin carecomposition comprises an effective amount of a first compositioncomprising a combination of a camellia serum fraction and a feverfewserum fraction, or a second composition comprising a combination of aparsley serum fraction and a kelp serum fraction; and a dermatologicallyacceptable carrier. According to another embodiment, a method forreducing the appearance of skin hypopigmentation is provided. The methodcomprises topically applying a skin care composition to a skin surface,wherein the skin care composition is applied for a period of timesufficient to stimulate one or more steps in melanogenesis (melaninsynthesis). The skin care composition comprises an effective amount of afirst composition comprising a combination of a camellia serum fractionand a feverfew serum fraction, or a second composition comprising acombination of a parsley serum fraction and a kelp serum fraction; and adermatologically acceptable carrier.

According to another embodiment, a method for reducing the appearance ofskin hypopigmentation is provided. The method comprises topicallyapplying a skin care composition to a skin surface, wherein the skincare composition is applied for a period of time sufficient to stimulateone or more steps in melanogenesis (melanin synthesis). The skin carecomposition comprises an effective amount of a combination of a camelliaserum fraction, a feverfew serum fraction, a parsley serum fraction, anda kelp serum fraction; and a dermatologically acceptable carrier.

According to one aspect of the present invention, a method is providedthat comprises topically applying the skin care composition comprisingan effective amount of a synergistic combination of skin tone agents toa skin surface for the purpose of improving the appearance of skin tone.

In response to the technical problems identified in the background, thepresent invention may take other forms. Further forms of the presentinvention will be appreciated in the detailed description that follows.

BRIEF DESCRIPTION OF THE DRAWINGS

It is believed that the present invention will be better understood fromthe following description taken in conjunction with the accompanyingdrawings. The referenced drawings are not to be construed as limitingthe scope of the present invention.

FIG. 1 is a bar graph showing melanin synthesis activation when kelp andparsley serum fractions are evaluated separately and as varied blends ina melanocyte assay.

FIG. 2 is another bar graph showing melanin synthesis activation whenkelp and parsley serum fractions are evaluated separately and as a 50:50blend in a melanocyte assay.

FIG. 3 is a bar graph showing melanin synthesis activation when camelliaand feverfew serum fractions are evaluated separately and as variedblends in a melanocyte assay.

FIG. 4 is another bar graph showing melanin synthesis activation whencamellia and feverfew serum fractions are evaluated separately and as20:80 blend in a melanocyte assay.

FIG. 5 is a bar graph showing inhibition of interleukin-1receptor-associated kinase (IRAK-4) when kelp and parsley serumfractions are evaluated separately and as varied blends in an ADP-Glo™assay.

FIG. 6 is a bar graph showing inhibition of IRAK-4 when camellia andfeverfew serum fractions are evaluated separately and as varied blendsin an ADP-Glo™ assay.

FIG. 7 is a table showing the identity of the serum fractions and blendsused herein.

DETAILED DESCRIPTION OF THE INVENTION

All percentages and ratios used herein are by weight of the totalcomposition and all measurements made are at 25° C., unless otherwisedesignated. All numeric ranges are inclusive of narrower ranges;delineated upper and lower range limits are interchangeable to createfurther ranges not explicitly delineated.

The compositions of the present invention can comprise, consistessentially of, or consist of, the essential components as well asoptional ingredients described herein. As used herein, “consistingessentially of” means that the composition or component may includeadditional ingredients, but only if the additional ingredients do notmaterially alter the basic and novel characteristics of the claimedcompositions or methods.

The term “apply” or “application” as used in reference to a composition,means to apply or spread the compositions of the present invention ontoa mammalian skin surface such as the epidermis.

The term “dermatologically acceptable” as used herein means that thecompositions or components described are suitable for use in contactwith human skin tissue without undue toxicity, incompatibility,instability, allergic response, and the like.

The term “effective amount” as used herein means an amount of a compoundor composition sufficient to significantly induce a positive benefit.

The term “post-inflammatory hypopigmentation” as used herein refers toan acute to chronic decrease in pigmentation as a response to atransient inflammatory event.

The term “hypopigmented spot” as used herein refers to a defined area ofskin wherein the pigmentation is less than that of an adjacent area ofskin due to localized and chronic or systemic underproduction ofmelanin.

The term “skin tone agent” as used herein refers to an agent thatregulates melanin production signals, synthesis of melanin, systemictransfer of melanin between the melanocyte and the keratinocyte, and/ormelanin degradation. Skin tone agents can improve the appearance ofuneven skin tone by acting as a pigmentation enhancing cosmetic agent.

The term “skin tone” as used herein refers to the overall appearance ofmelanin in the skin caused by the systemic, rather than transient,synthesis of melanin. Skin tone is typically characterized over a largerarea of the skin. The area ideally may be than 100 mm², but larger areasare envisioned such as the entirety of the facial skin or any of thefacial skin surfaces. Skin tone can be measured by image analysis. Forexample, overall lightness can be measured by L* coordinate in L*a*b*color space (International Commission on Illumination). Chromophoremapping such as melanin mapping and melanin concentration may be used asan indicator of overall skin tone. Mean melanin may be calculated fromthe chromophore map data. Additionally, skin tone evenness can bedetermined by melanin evenness which also may be calculated from thechromophore map data. Suitable chromophore mapping techniques arediscussed in the example below.

The term “facial skin surface” as used herein refers to one or more offorehead, periorbital, cheek, perioral, chin, and nose skin surfaces.

As used herein, “exogenous solvent” means any solvent that is notinherently present in the plant material, but is placed in contact withthe plant material for the purpose of separating (e.g., extracting)compounds from the plant material.

The term “serum fraction,” as used herein, means a composition producedby a general method comprising the steps of: (a) grinding and pressingof fresh plant matter and (b) separating a liquid fraction from a cellwall fraction to obtain fresh cell juice, wherein no exogenous liquid isadded prior or during said separating; (c) filtering the fresh celljuice to obtain a first filtrate; and (d) fractionating the firstfiltrate to obtain the serum fraction.

I. Skin Care Compositions

An embodiment of the present invention relates to various compositionsand, more specifically, to compositions for application to a skinsurface, which does do not include a scalp surface. The skin carecompositions comprise an effective amount of skin tone agents, which areeffective for providing evenly toned skin, and a dermatologicallyacceptable carrier. The compositions may be in a wide variety of productforms that include, but are not limited to, solutions, suspensions,lotions, creams, gels, toners, sticks, pencil, sprays, aerosols,ointments, cleansing liquid washes and solid bars, skin conditioners,pastes, foams, powders, mousses, shaving creams, wipes, strips, patches,electrically-powered patches, wound dressing and adhesive bandages,hydrogels, film-forming products, facial and skin masks (with andwithout insoluble sheet), make-up such as foundations, and eye shadows,and the like. The composition form may follow from the particulardermatologically acceptable carrier chosen.

A. Skin Tone Agents

The skin care compositions in accordance to embodiments of the presentinvention comprise an effective amount of a combination of skin toneagents which are blends of serum fractions. Accordingly, in anembodiment, the skin care composition comprises a first combination of acamellia serum fraction and a feverfew serum fraction; or a secondcombination of a parsley serum fraction and a kelp serum fraction.According to another embodiment the skin care composition comprises thefirst and the second combinations.

The camellia, feverfew, or parsley fractions each consist essentially ofthe flower, leaf, and/or stem serum fractions obtained from plantsbelonging to camellia sinensis, chrysanthemum parthenium, orpetroselinum crispum, respectively, and the kelp serum fraction consistsessentially of a serum fraction from a photosynthetic organismmacrocystis pyrifera, which is commonly known as brown algae. Forpurposes of simplifying the discussion herein, brown algae should beunderstood to be inclusive to “plant” or “plants” or “biomass” whenreferencing the source of a serum fraction. Exemplary serum fractionsused herein are provided by Akzo Nobel Surface Chemistry LLC, Chicago,Ill. Exemplary serum fraction preparation methods are set forth in U.S.Pat. No. 7,473,435 (e.g., for camellia), U.S. Pat. No. 7,537,791 (e.g.,for parthenolide free bioactive ingredients from feverfew (Tanacetumparthenium)), and U.S. Patent Application Publication No. 2011/0110872(e.g., for kelp), which are incorporated herein by reference in theirentirety. The serum fractions may be blended to form a combination ofserum fractions that surprisingly produce synergistic effects, asdiscussed further below.

The general method for preparing a serum fraction comprises the stepsof: grinding and pressing of clean, fresh plant matter; separating aliquid fraction from a cell wall fraction to obtain fresh cell juice,wherein no exogenous liquid is added prior or during said separating;filtering the fresh cell juice to obtain a first filtrate; andfractionating the first filtrate to obtain the serum fraction for useherein. Fractionating may include one or more of the following steps:adjusting pH; heating such as microwaving; filtering and/or centrifugingto remove chlorophyll, pigments, and/or proteins from said firstfiltrate to form the serum fraction. The method may further includestabilizing to serum fraction. Stabilizing may include addingpreservatives and incubating the mixture until complete solubilizationof the preservative is achieved. Exemplary preservatives include one ormore of potassium sorbate, sodium benzoate, sodium methyl paraben,and/or citric acid.

The resulting combinations or blends of serum fractions have superiorbioactivity versus traditionally prepared plant extracts. Unliketraditional extracts, the serum fraction is prepared from fresh plantcell juice that has been mechanically separated from the rest of thefresh plant material. Importantly, no exogenous solvent (e.g., water,hexane, acetone, ethanol) is added during the juice separation process.The resulting cell juice contains the full spectrum of compounds foundin fresh plant matter, thus the resulting serum fractions contain a muchbroader range of active compounds than do traditional plant extracts,which contain only the narrow range of compounds that can be separatedwith a particular solvent.

Furthermore, using fresh plants maintains the integrity of the bioactivecomponents inherently present in the fresh plant matter. Traditionalplant extracts are not prepared from fresh plant matter, but rather fromdried plant material, which has undergone degradation due todehydration. During dehydration, the cell walls are compromised, causingthe degredation of compounds through mechanisms such as hydrolysis,oxidation, polymerization, Maillard reactions, and isomerization. Whenthe dried leaves are extracted, the resulting extract thus containsthese degradation products that were not originally present in the freshplant matter. Accordingly, the composition of the resulting dry leafextract greatly differs from that of fresh juice and the resulting serumfraction.

An exemplary preparation of a camellia sinensis serum fraction isdescribed in U.S. Pat. No. 7,473,435, and is summarized below. The serumfraction from camellia sinensis plants can be prepared by a methodcomprising the steps of (1) biomass preparation; (2) grinding,maceration, and pressing of plant biomass; (3) separation of a membranefraction from the cell juice to provide a cell juice supernatant; (4)separation of a cytoplasm fraction from the cell juice supernatant; and(5) isolation of the serum fraction.

(1) Biomass Preparation: Sufficient amounts of fresh camellia (camelliasinensis) plant biomass (only top tender young leaf tissue with buds)are harvested to yield approximately 100 kg of dry matter. The level ofdry matter in the fresh biomass is calculated to be 21.70 wt %,requiring harvesting of approximately 461 kg of fresh plant biomass toyield 100 kg of dry matter. Care is taken to preserve the inherentmoisture content of the plant biomass and to avoid wilting due tomoisture loss. The harvesting is conducted in such a manner as to avoidor minimize chopping, mashing, and crushing of the collected biomass toavoid the disruption of the leaf cell structure, which can induceendogenous enzymatic reactions catalized by phenol-oxidase andperoxidase. Because these reactions are intensified with the time ofoxidation, all steps are completed in the shortest possible period oftime. For example, the harvested biomass is delivered for processing notmore than 10 minutes after cutting. This is done to minimize exposure ofthe plant biomass to sun, high temperature, and other negativeenvironmental factors. A washing step is performed to remove soilparticles and other debris from the plants prior to further processing.This washing is accomplished by washing the harvested plants for ≤5minutes in ≤1 kg/cm² water pressure. The residual water wash does notcontain any green or brown pigments, indicating proper water pressureand washing duration. The excess water is removed from the washed plantbiomass.

(2) Grinding, Maceration, and Pressing of Plant Biomass: Afterharvesting, collecting, and washing the plant biomass, the plants thenundergo grinding, maceration, and pressing to extract the intracellularcontent (i.e., the plant cell juice) and to separate the plant celljuice from the fiber-enriched cell walls fraction (cell walls fraction).A hammer mill (Model VS 35, Vincent Corporation, Fla.) having 10 HPengine and set of screens may be used to grind the biomass to yieldplant tissue particles of suitably small size in a shortest amount oftime and without significant increase of biomass temperature. The hammermill can be set to produce the maximum size of macerated plant particlesof ≤0.5 centimeters during ≤10 seconds of treatment. The biomasstemperature is increased only ≤5° C. A horizontal continuous screw press(Compact Press “CP-6”, Vincent Corporation, Fla.) is immediately used toextract the plant cell juice from the plant. The pressure on the cone ofthe screw press is maintained at a level of 24 kg/cm², with a screwspeed of 12 rpm and only a temperature increase of ≤5° C. This treatmentcan yield about 276 kg of plant cell juice having dry mater level ofabout 8.5 wt %.

(3) Separation of the Membrane Fraction from the Cell Juice: The initialplant cell juice having dry matter level of about 8.5 wt % containssmall fiber particles, which can be removed by filtration through fourlayers of nylon fabric or by using low-speed centrifugation biomass. Thefiltered plant cell juice is exposed to microwave treatment using atemperature probe control. This treatment continues until thetemperature of the cell juice reached 60° C. Once coagulation isinduced, the treated cell juice is immediately cooled to 40° C.Separation of the membrane fraction from the coagulated cell juice isachieved using centrifugation at greater than or equal to 3,000 g forgreater than or equal to 20 minutes. This yields a membrane fraction(precipitate) and a cell juice supernatant, which contains a cytoplasmfraction and a cell serum fraction (which contains low molecular weightsoluble components). The cell juice supernatant is used for furtherprocessing to yield a serum fraction.

(4) Separation of the Cytoplasm Fraction from the Cell JuiceSupernatant: In order to separate out the cytoplasm fraction, the celljuice supernatant is subjected to isoelectric precipitation.Precipitation of the cytoplasm fraction is induced using a titrationmethod utilizing 5.0N Hydrochloric Acid (HCl) to bring the pH of thecell juice supernatant to about 4. The separation of precipitatedcytoplasm fraction, which may have a dry matter level of about 14.5 wt%, from supernatant is achieved by centrifugation at greater than orequal to 3,000 g (where g is the relative centrifugal force) for greaterthan or equal to 20 minutes.

(5) Isolation of the Serum Fraction: After separation of the cytoplasmfraction, the supernatant contains suspended particles. In order toseparate out these particles, the supernatant is centrifuged at greaterthan or equal to 7,500 g for greater than or equal to 30 minutes. Thetransparent supernatant is filtered through a filter having 0.8micrometer pores. This filtrate (camellia sinensis serum fraction) canhave a dry matter level of about 5.7 wt %, which is based on the recitedprocess using no exogenous extraction solvents.

It can be appreciated that the dry matter level of the serum fractioncan vary depending on a variety of factors such as the moisture contentof the plant biomass, which itself may have variations based on seasonaland/or geographical source. Accordingly, in one embodiment the drymatter level in the camellia serum fraction can range from about 0.1 wt% to about 25 wt %, from about 1 wt % to about 15 wt %, from about 2 wt% to about 10 wt %, or from about 3 wt % to about 9 wt %, for example.In one embodiment, the dry matter level in the feverfew fraction canrange from about 0.1 wt % to about 25 wt %, from about 1 wt % to about15 wt %, from about 2 wt % to about 10 wt %, or from about 3 wt % toabout 9 wt %, for example. In one embodiment, the dry matter level inthe kelp fraction can range from about 0.1 wt % to about 25 wt %, fromabout 1 wt % to about 15 wt %, from about 2 wt % to about 10 wt %, orfrom about 3 wt % to about 9 wt %, for example. In one embodiment, thedry matter level in the parsley fraction can range from about 0.1 wt %to about 25 wt %, from about 1 wt % to about 15 wt %, from about 2 wt %to about 10 wt %, or from about 3 wt % to about 9 wt %, for example. Inanother embodiment, the dry matter level in the blend of the kelp andparsley serum fractions and/or the camellia and feverfew serum fractionscan range from about 0.1 wt % to about 25 wt %, from about 1 wt % toabout 15 wt %, from about 2 wt % to about 10 wt %, or from about 3 wt %to about 9 wt %, for example.

Additionally, the serum fractions can be further characterized withrespect to the content of specified compounds or classes of compounds inthe serum fraction and/or the dry matter, or by the absence of othercompounds or class of compounds. For example, in one embodiment, thecamellia serum fraction can have a total catechin content of betweenabout 8.0 and about 20.0 milligrams per gram of dry matter, particularlybetween about 10.0 and about 18.0 milligrams per gram of dry matter, andmore particularly between about 12.0 and about 16.0 milligrams per gramof dry matter. In another example, the feverfew serum fraction is eitherfree of, or substantially free of, α-unsaturated γ-lactones, such asparthenolide.

Similar, analysis and characterization can applied to the feverfew (seee.g., U.S. Pat. No. 7,537,791), kelp (see e.g., U.S. Patent ApplicationPublication No. 2011/0110872), and parsley serum fractions.

In some instances, a serum fraction cannot be used as an activeingredient of topical products due to a lack of stability anddeterioration of color and odor. A refinement of the serum fraction mayinvolve the following steps: heat treatment, cooling, filtration, andstabilization. Refinement can be performed immediately after separationof the serum fraction from the cytoplasm fraction. For example, thecamellia serum fraction is exposed to microwave treatment using atemperature probe control. This treatment continues until thetemperature of the serum fraction reaches 99° C. (90° C. is required aswas previously described in U.S. Pat. No. 7,537,791, which is herebyincorporated by reference in its entirety). Once coagulation is induced,the treated serum fraction is immediately cooled to 10° C. Thecoagulated serum fraction is vacuum filtrated through filter havingporous 0.8 micrometer (double layers of Whatman No. 2 filters may alsobe used as described in U.S. Pat. No. 7,537,791). The precipitate may bediscarded and the resulting serum fraction filtrate can undergo furtherprocessing for stabilization. Stabilization of the serum fractionfiltrate is achieved by adding preservatives (no exogenous anti-oxidantis required as was previously described in U.S. Pat. No. 7,537,791) andincubating the mixture until complete solubilization is achieved. Thepreservatives useful for stabilization of the serum fraction filtrateinclude the following: 0.1% potassium sorbate, 0.1% sodium benzoate,0.1% sodium methyl paraben, and/or 0.1% citric acid. The recommendedstorage conditions for the camellia serum fraction includes storage in aclosed container protected from light at a temperature of between 15° C.and 25° C.

Similar processing of chrysanthemum parthenium, petroselinum crispum, ormacrocystis pyrifera can yield the serum fractions of feverfew, parsley,or kelp, respectively.

In some embodiments, the skin care composition comprises an effectiveamount of skin tone agents that comprises a first combination of acamellia serum fraction and a feverfew serum fraction, the camelliaserum fraction being present in an amount of from about 0.001 wt % toabout 15 wt %, alternatively from about 0.002 wt % to about 10 wt %,alternately from about 0.025 wt % to about 10 wt %, in other embodimentsfrom about 0.05 wt % to about 10 wt %, in others from about 0.05 wt % toabout 5 wt %, and in others from about 0.1 wt % to about 5 wt %; and thefeverfew serum fraction being present in an amount from about 0.001 wt %to about 15 wt %, from about 0.002 wt % to about 10 wt %, from about0.025 wt % to about 10 wt %, from about 0.05 wt % to about 10 wt %, fromabout 0.05 wt % to about 5 wt %, or from about 0.1 wt % to about 5 wt %,wherein the wt % is based on the weight of the skin care composition.

According to another embodiment, weight ratio of the camellia serumfraction to the feverfew serum fraction ranges from about 10:90 to about90:10, from about 10:90 to about 50:50; from about 80:20 to about 20:80,from about 30:70 to about 70:30, from about 40:60 to about 60:40. Forexample, the weight ratio of the camellia serum fraction to feverfewserum fraction may be about 90:10, about 80:20, about 70:30, about60:40, about 50:50, about 40:60, about 30:70, about 20:80, or about10:90.

In another embodiment, the skin care composition comprises an effectiveamount of a second combination of skin tone agents that comprises aparsley serum fraction and a kelp serum fraction, the parsley serumfraction being present in an amount from about 0.001 wt % to about 15 wt%, from about 0.002 wt % to about 10 wt %, from about 0.025 wt % toabout 10 wt %, from about 0.05 wt % to about 10 wt %, from about 0.05 wt% to about 5 wt %, or from about 0.1 wt % to about 5 wt %; and the kelpserum fraction being present in an amount from about 0.001 wt % to about15 wt %, from about 0.002 wt % to about 10 wt %, from about 0.025 wt %to about 10 wt %, from about 0.05 wt % to about 10 wt %, from about 0.05wt % to about 5 wt %, or from about 0.1 wt % to about 5 wt %, whereinthe wt % is based on the weight of the skin care composition.

According to another embodiment, weight ratio of the parsley serumfraction to the kelp serum fraction ranges from about 10:90 to about90:10, from about 10:90 to about 50:50; from about 80:20 to about 20:80,from about 30:70 to about 70:30, from about 40:60 to about 60:40. Forexample, the weight ratio of the parsley serum fraction to kelp serumfraction may be about 90:10, about 80:20, about 70:30, about 60:40,about 50:50, about 40:60, about 30:70, about 20:80, or about 10:90.

B. Dermatologically Acceptable Carrier

The compositions of the present invention may also comprise adermatologically acceptable carrier (which may be referred to as“carrier”) for the composition. The phrase “dermatologically acceptablecarrier”, as used herein, means that the carrier is suitable for topicalapplication to the keratinous tissue, has good aesthetic properties, iscompatible with the skin tone agents in the composition, and will notcause any unreasonable safety or toxicity concerns. A suitable carrieris selected to yield a desired product form. Furthermore, the solubilityor dispersibility of the components may dictate the form and characterof the carrier. In one embodiment, the carrier is present at a level offrom about 50 wt % to about 99 wt %, about 60 wt % to about 98 wt %,about 70 wt % to about 98 wt %, or, alternatively, from about 80 wt % toabout 95 wt %, by weight of the composition. In another embodiment, thecarrier is present in the skin care composition at a level of from about25 wt % to about 50 wt %, which may be diluted upon application with asuitable carrier diluent.

The carrier can be in a wide variety of forms. Non-limiting examplesinclude simple solutions (e.g., aqueous, organic solvent, or oil based),emulsions, and solid forms (e.g., gels, sticks, flowable solids, oramorphous materials). In certain embodiments, the dermatologicallyacceptable carrier is in the form of an emulsion. Emulsion may begenerally classified as having a continuous aqueous phase (e.g.,oil-in-water and water-in-oil-in-water) or a continuous oil phase (e.g.,water-in-oil and oil-in-water-in-oil). The oil phase of the presentinvention may comprise silicone oils, non-silicone oils such ashydrocarbon oils, esters, ethers, and the like, and mixtures thereof.

The aqueous phase comprises water, such as demineralized or distilledwater, for example. Other acceptable carriers that may be used in theaqueous carrier include, but are not limited to alcohol compounds, suchas ethanol. According to one embodiment, the composition comprisesalcohol, dipropylene glycol, and/or water.

The skin care compositions have a pH ranging from about 3.0 to about 10,which may be measured by taking a direct pH measurement using a standardhydrogen electrode of the composition at 25° C. Accordingly, the pH ofthe skin care composition may be within the range from about 6 to about9, for example.

Emulsions may further comprise an emulsifier. The composition maycomprise any suitable percentage of emulsifier to sufficiently emulsifythe carrier. Suitable weight ranges include from about 0.1 wt % to about10 wt % or about 0.2 wt % to about 5 wt % of an emulsifier, based on theweight of the composition. Emulsifiers may be nonionic, anionic orcationic. Suitable emulsifiers are disclosed in, for example, U.S. Pat.No. 3,755,560, U.S. Pat. No. 4,421,769, and McCutcheon's Detergents andEmulsifiers, North American Edition, pages 317-324 (1986), which areincorporated herein by reference in their entirety. Suitable emulsionsmay have a wide range of viscosities, depending on the desired productform.

C. Optional Ingredients

According to embodiments of the present invention, the skin carecompositions can also additionally comprise suitable optionalingredients as desired. For example, the composition can optionallyinclude other active or inactive ingredients, provided that they do notunacceptably alter the benefits of the skin care composition. Whenpresent, compositions of the present invention may contain from about0.0001% to about 50%; from about 0.001% to about 20%; or, alternately,from about 0.01% to about 10%, by weight of the composition, of theoptional components. The amounts listed herein are only to be used as aguide, as the optimum amount of the optional components used in acomposition will depend on the specific active selected since theirpotency does vary considerably. Hence, the amount of some optionalcomponents useful in the present invention may be outside the rangeslisted herein.

The optional components, when incorporated into the composition, shouldbe suitable for use in contact with human skin tissue without unduetoxicity, incompatibility, instability, allergic response, and the like.The compositions of the present invention may include optionalcomponents such as anti-acne actives, desquamation actives,anti-cellulite agents, chelating agents, flavonoids, tanning active,non-vitamin antioxidants and radical scavengers, skin growth regulators,anti-wrinkle actives, anti-atrophy actives, minerals, phytosterolsand/or plant hormones, N-acyl amino acid compounds, antimicrobial orantifungal actives, and other useful skin care actives, which aredescribed in further detail in U.S. application publication No.US2006/0275237A1 and US2004/0175347A1.

According to yet another embodiment, the skin care composition mayfurther include one or more additional skin care actives. Accordingly,non-limiting examples of additional skin care actives include flavonoidssuch as apigenin and luteolin, indole compounds, xanthine compounds,vitamin B₃ compounds, panthenol compounds, and derivatives thereof.

1. Flavonoids

The compositions of embodiments of the present invention may optionallycontain a flavonoid compound. Flavonoids are broadly disclosed in U.S.Pat. Nos. 5,686,082 and 5,686,367, both of which are herein incorporatedby reference. Flavonoids suitable for use in embodiments of the presentinvention are flavanones selected from unsubstituted flavanones,mono-substituted flavanones, and mixtures thereof; chalcones selectedfrom unsubstituted chalcones, mono-substituted chalcones, di-substitutedchalcones, tri-substituted chalcones, and mixtures thereof; flavonesselected from unsubstituted flavones, mono-substituted flavones,di-substituted flavones, and mixtures thereof; one or more isoflavones;coumarins selected from unsubstituted coumarins, mono-substitutedcoumarins, di-substituted coumarins, and mixtures thereof; chromonesselected from unsubstituted chromones, mono-substituted chromones,di-substituted chromones, and mixtures thereof; one or more dicoumarols;one or more chromanones; one or more chromanols; isomers (e.g.,cis/trans isomers) thereof; and mixtures thereof. By the term“substituted” as used herein means flavonoids wherein one or morehydrogen atom of the flavonoid has been independently replaced withhydroxyl, C1-C8 alkyl, C1-C4 alkoxyl, 0-glycoside, and the like or amixture of these substituents.

Examples of suitable flavonoids include, but are not limited to,unsubstituted flavanone, mono-hydroxy flavanones (e.g., 2′-hydroxyflavanone, 6-hydroxy flavanone, 7-hydroxy flavanone, etc.), mono-alkoxyflavanones (e.g., 5-methoxy flavanone, 6-methoxy flavanone, 7-methoxyflavanone, 4′-methoxy flavanone, etc.), unsubstituted chalcone(especially unsubstituted trans-chalcone), mono-hydroxy chalcones (e.g.,2′-hydroxy chalcone, 4′-hydroxy chalcone, etc.), di-hydroxy chalcones(e.g., 2′,4-dihydroxy chalcone, 2′,4′-dihydroxy chalcone, 2,2′-dihydroxychalcone, 2′,3-dihydroxy chalcone, 2′,5′-dihydroxy chalcone, etc.), andtri-hydroxy chalcones (e.g., 2′,3′,4′-trihydroxy chalcone,4,2′,4′-trihydroxy chalcone, 2,2′,4′-trihydroxy chalcone, etc.),unsubstituted flavone, 7,2′-dihydroxy flavone, 3′,4′-dihydroxynaphthoflavone, 4′-hydroxy flavone, 5,6-benzoflavone, and7,8-benzoflavone, unsubstituted isoflavone, daidzein (7,4′-dihydroxy isoflavone), 5,7-dihydroxy-4′-methoxy isoflavone, soy isoflavones (amixture extracted from soy), unsubstituted coumarin, 4-hydroxy coumarin,7-hydroxy coumarin, 6-hydroxy-4-methyl coumarin, unsubstituted chromone,3-formyl chromone, 3-formyl-6-isopropyl chromone, unsubstituteddicoumarol, unsubstituted chromanone, unsubstituted chromanol, andmixtures thereof.

In an embodiment, the flavonoid compound is an unsubstituted flavanone,methoxy flavanones, unsubstituted chalcone, 2′,4-dihydroxy chalcone, andmixtures thereof. For example, in another embodiment, are unsubstitutedflavanone, unsubstituted chalcone (e.g., the trans isomer), and mixturesthereof.

Flavonoids can be synthetic materials or obtained as extracts fromnatural sources (e.g., plants). The naturally sourced material can alsofurther be derivatized (e.g., an ester or ether derivative preparedfollowing extraction from a natural source). Flavonoid compounds usefulherein are commercially available from a number of sources, e.g.,Indofine Chemical Company, Inc. (Somerville, N.J.), Steraloids, Inc.(Wilton, N.H.), and Aldrich Chemical Company, Inc. (Milwaukee, Wis.).

Mixtures of the above flavonoid compounds may also be used.

When included in the skin care composition, the herein describedflavonoid compounds can be present at concentrations of from about 0.01wt % to about 20 wt %, of from about 0.1 wt % to about 10 wt %, or fromabout 0.5 wt % to about 5 wt %, wherein the wt % is based on the totalweight of the skin care composition.

2. Indole Compounds

The skin care compositions can further include an indole compound. Asused herein, “indole compound” means one or more indoles, derivativesthereof, mixtures thereof, or salts thereof. Accordingly, thecomposition may include from about 0.1 wt % to about 10 wt % of theindole compound, from about 0.5 wt % to about 5 wt % of the indolecompound, or from about 1 wt % to about 2 wt % of the indole compound,for example, wherein the wt % is based on the total weight of the skincare composition.

3. Xanthine Compounds

The skin care compositions can further include a xanthine compound. Asused herein, “xanthine compound” means one or more xanthines,derivatives thereof, and mixtures thereof. Xanthine compounds that canbe useful herein include, but are not limited to, caffeine, xanthine,1-methylxanthine, theophylline, theobromine, derivatives thereof, andmixtures thereof. Accordingly, the composition may include from about0.1 wt % to about 10 wt % of the xanthine compound, from about 0.5 wt %to about 5 wt % of the xanthine compound, or from about 1 wt % to about2 wt % of the xanthine compound, for example, wherein the wt % is basedon the total weight of the skin care composition. For example, the skincare composition may further include about 0.75 wt % of caffeine.

4. Vitamin B₃ Compounds

The skin care compositions can further include a vitamin B₃ compound. Asused herein, “vitamin B₃ compound” means nicotinic acid, niacinamide,nicotinyl alcohol, derivatives thereof, and mixtures thereof. Thevitamin B₃ compound may be included as the substantially pure material,or as an extract obtained by suitable physical and/or chemical isolationfrom natural (e.g., plant) sources. Accordingly, the composition mayinclude from about 0.1 wt % to about 25 wt % of the vitamin B₃ compound;from about 0.5 wt % to about 15 wt % of the vitamin B₃ compound; or fromabout 3.5 wt % to about 7.5 wt % of the vitamin B₃ compound, forexample, wherein the wt % is based on the total weight of the skin carecomposition. For example, the skin care composition may further includeabout 2.5 wt % of vitamin B₃.

5. Panthenol Compounds

The skin care compositions can further comprise a panthenol compound. Asused herein, the term “panthenol compound” includes panthenol, one ormore pantothenic acid derivatives, and mixtures thereof. Non-limitingexamples of panthenol compounds include D-panthenol([R]-2,4-dihydroxy-N-[3-hydroxypropyl)]-3,3-dimethylbutamide),D,L-panthenol, pantothenic acids and their salts (e.g., the calciumsalt), panthenyl triacetate, royal jelly, panthetine, pantotheine,panthenyl ethyl ether, pangamic acid, pantoyl lactose, Vitamin Bcomplex, or mixtures thereof. Accordingly, the composition may includefrom about 0.01 wt % to about 5 wt % of the panthenol compound; fromabout 0.03 wt % to about 3 wt % of the panthenol compound; from about0.05 wt % to about 2 wt % of the panthenol compound; or from about 0.1wt % to about 1 wt % of the panthenol compound, for example, wherein thewt % is based on the total weight of the skin care composition. Forexample, the skin care composition may further include about 0.15 wt %of panthenol.

6. Additional Skin Tone Agent

In some embodiments, it may be desirable to include one or moreadditional skin tone agents in the composition. The additional skin toneagents can be included to further improve overall skin tone. Whenpresent, the compositions of the present invention contain up to about50%, 40%, 30%, 20%, 10%, 5%, or 3%, by weight of the composition, of theskin tone agent. When present, the compositions of the present inventioncontain at least about 0.001%, 0.01%, 0.1%, 0.2%, 0.5%, or 1%, by weightof the composition, of the additional skin tone agent. Suitable rangesinclude any combination of the lower and upper limits including suitableranges from about 0.1% to about 50%; from about 0.2% to about 20%; orfrom about 1% to about 10%, by weight of the composition, of the skintone agent. The amounts listed herein are only to be used as a guide, asthe optimum amount of the skin tone agent will depend on the specificactive selected since their potency does vary considerably.

Suitable additional skin tone agents include, but are not limited to,sugar amines, vitamin B3 compounds, arbutin, deoxyarbutin,1,3-dihydroxy-4-alkylbenzene such as hexylresorcinol, sucrosedilaurante, bakuchoil (4-[(1E,3S)-3-ethenyl-3,7-dimethyl-1,6octadienyl]phenol or monterpene phenol), pyrenoine (available fromBiotech Marine, France), panicum miliaceum seed extract, arlatone dioicacid, cinnamic acid, ferulic acid, achromaxyl, methyl nicotinamide, oilsoluble licorice extract, folic acid, undecylenic acid (i.e., undecenoicacid), zinc undecylenate, thiamine (Vitamin B1) and its hydrochloride,L-tryptophan, helianthus annuus (sunflower) and vitis vinifera (grape)leaf extract, carnosine (i.e., dragosine), methyl gentisate,1,2-hexandiol and 1,2-octandiol (i.e., combination sold as Symdiol 68 bySymrise AG, Germany), inositol, decylenoylphenylalanine (e.g., soldunder the tradename Sepiwhite by Seppic, France), koijic acid,hexamidine compounds, salicylic acid, and retinoids including retinoland retinol propionate.

In certain embodiments, the additional skin tone agent is selected fromvitamin B3 compounds, sugar amines, hexamidine compounds, salicylicacid, 1,3-dihydroxy-4-alkylbenzene such as hexylresorcinol, andretinoids. As used herein, “vitamin B₃ compound” means a compound havingthe formula:

wherein R is —CONH₂ (i.e., niacinamide), —COOH (i.e., nicotinic acid) or—CH₂OH (i.e., nicotinyl alcohol); derivatives thereof; and salts of anyof the foregoing. As used herein, “sugar amine” includes isomers andtautomers of such and its salts (e.g., HCl salt) and its derivatives.Examples of sugar amines include glucosamine, N-acetyl glucosamine,mannosamine, N-acetyl mannosamine, galactosamine, N-acetylgalactosamine, their isomers (e.g., stereoisomers), and their salts(e.g., HCl salt). As used herein, “hexaminide compound” means a compoundhaving the formula:

wherein R¹ and R² are optional or are organic acids (e.g., sulfonicacids, etc.). In one embodiment, hexamidine compound is hexamidinediisethionate.

7. Additional Anti-Inflammatory Agents Hypopigmentation may result fromskin inflammation. Transient inflammatory events triggeringhypopigmentation and, more specifically, post-inflammatoryhypopigmentation include, but are not limited to, acne lesions, ingrownskins, scratches, insect bites, surfactant damage, allergens, andshort-term UV exposure. Inflammation induced hypopigmentation includingpost-inflammatory hypopigmentation may be managed by the skin carecompositions provide herein, insofar as the blend of the camellia serumfraction and the feverfew serum fraction, and the blend of the kelpserum fraction and the parsley serum fraction inhibit inflammation, asindicated by the inhibition of interleukin-1 receptor-associated kinase(IRAK-4). However, by incorporating into the compositions of the presentinvention an additional anti-inflammatory agent, enhanced or synergisticbenefits may be achieved. When present, the compositions of the presentinvention contain up to about 20%, 10%, 5%, 3%, or 1% by weight of thecomposition, of the additional anti-inflammatory agent. When present,the compositions of the embodiments of the present invention contain atleast about 0.001%, 0.01%, 0.1%, 0.2%, 0.3%, 0.5%, or 1%, by weight ofthe composition, of the additional anti-inflammatory agent. Suitableranges include any combination of the lower and upper limits. Suitableadditional anti-inflammatory agents include, but are not limited tononsteroidal anti-inflammatory agents (NSAIDS including but not limitedto ibuprofen, naproxen, flufenamic acid, etofenamate, aspirin, mefenamicacid, meclofenamic acid, piroxicam and felbinac), glycyrrhizic acid(also known as glycyrrhizin, glycyrrhixinic acid, and glycyrrhetinicacid glycoside) and salts such as dipotassium glycyrrhizate,glycyrrhetenic acid, licorice extracts, bisabolol (e.g., alphabisabolol), manjistha (extracted from plants in the genus Rubia,particularly Rubia cordifolia), and guggal (extracted from plants in thegenus Commiphora, particularly Commiphora mukul), kola extract,chamomile, red clover extract, and sea whip extract (extracts from plantin the order Gorgonacea), derivatives of any of the foregoing, andmixtures thereof.

8. Sunscreen Actives

The compositions of the subject invention may comprise one or moresunscreen actives (or sunscreen agents) and/or ultraviolet lightabsorbers. Herein, “sunscreen active” collectively includes, sunscreenactives, sunscreen agents, and/or ultraviolet light absorbers. Sunscreenactives include both sunscreen agents and physical sunblocks. Sunscreenactives may be organic or inorganic. Examples of suitable sunscreenactives are disclosed in Personal Care Product Council's InternationalCosmetic Ingredient Dictionary and Handbook, Thirteenth Edition, as“sunscreen agents.” Particularly suitable sunscreen actives are2-ethylhexyl-p-methoxycinnamate (commercially available as PARSOL™ MCX),4,4′-t-butyl methoxydibenzoyl-methane (commercially available as PARSOL™1789), 2-hydroxy-4-methoxybenzophenone, octyldimethyl-p-aminobenzoicacid, digalloyltrioleate, 2,2-dihydroxy-4-methoxybenzophenone,ethyl-4-(bis(hydroxypropyl))aminobenzoate,2-ethylhexyl-2-cyano-3,3-diphenylacrylate, 2-ethylhexyl-salicylate,glyceryl-p-aminobenzoate, 3,3,5-tri-methylcyclohexylsalicylate, menthylanthranilate, p-dimethyl-aminobenzoic acid or aminobenzoate,2-ethylhexyl-p-dimethyl-amino-benzoate, 2-phenylbenzimidazole-5-sulfonicacid, 2-(p-dimethylaminophenyl)-5-sulfonicbenzoxazoic acid, octocrylene,zinc oxide, benzylidene camphor and derivatives thereof, titaniumdioxide, and mixtures thereof.

In one embodiment, the composition may comprise from about 1% to about20%, and alternatively from about 2% to about 10% by weight of thecomposition, of the sunscreen active. Exact amounts will vary dependingupon the chosen sunscreen active and the desired Sun Protection Factor(SPF), which is within the knowledge of one of skilled in the art.

II. Methods of Treatment

Various methods of treatment, application, regulation, or improvementmay utilize the aforementioned compositions. In one embodiment, themethod includes the step of identifying a hypopigmented spot forimprovement by the composition. The hypopigmented spot may be identifiedby the user or a third party such as a dermatologist, cosmetician, orother caregiver. Identification may be done by visual inspection of theskin for hypopigmented spots in need of treatment based on size and/orcolor. Identification may also be done by commercially available imagingdevices such SIAscope® V (available from Astron Clinica, Ltd., UK) orthe VISIA® Complexion Analysis system (available from CanfieldScientific, Inc., Fairfield, N.J.). Both devices are capable ofcollecting images of the skin and identifying hypopigmented spots. Insome instances, the method comprises the step of identifying a pluralityof hypopigmented spots for treatment by the composition.

Identification of the hypopigmented spot may occur on any skin surfaceof the body. Skin surfaces of the most concern tend to be those nottypically covered by clothing such as facial skin surfaces, hand and armskin surfaces, foot and leg skin surfaces, and neck and chest skinsurfaces. In particular, identification of the hypopigmented spot may beon a facial skin surface including the forehead, perioral, chin,periorbital, nose, and/or cheek skin surfaces.

The method may comprise the step of applying the composition to ahypopigmented spot or spots, which may have been previously identified.Many regimens exist for the application of the composition to thehypopigmented spot. The composition may be applied at least once a day,twice a day, or on a more frequent daily basis, during a treatmentperiod. When applied twice daily, the first and second applications areseparated by at least 1 to about 12 hours. Typically, the compositionmay be applied in the morning and/or in the evening before bed.

The treatment period is ideally of sufficient time to provide animprovement in the hypopigmented spot. The improvement may be adetectable reduction in size of the hypopigmented spot, darkening of thehypopigmented spot, increase in melanin of the hypopigmented spot, orevening of the tone of skin. The treatment period may be at least about1 week. The treatment period may last about 4 weeks or about 8 weeks. Incertain embodiments, the treatment period will extend over multiplemonths (i.e., 3-12 months) or multiple years. In one embodiment thecomposition is applied to the hypopigmented spot(s) at least once a dayduring a treatment period of at least about 4 weeks or at least about 8weeks. In one embodiment the composition is applied to the hypopigmentedspot(s) twice a day during a treatment period of at least about 4 weeksor 8 weeks.

In still another embodiment, the method comprises applying the skin carecomposition according to a regimen, wherein said regimen comprises:

(a) cleansing the skin to form a cleansed skin surface;

(b) topically applying the skin care composition to said cleansed skinsurface.

The skin care composition may be used daily, weekly, or in a variety ofregimens. The skin care composition may be used more than once a day,such as at night and in the morning. The skin care composition may beused more than once per day on certain days or use only a few times perweek. The skin care composition may be used three times per day, twiceper day, once per day, six times per week, five times per week, fourtimes per week, three times per week, two times per week, or one timeper week. In some embodiments, the skin care composition is used four,five, six or seven times per week.

The step of applying the composition to the hypopigmented spot may beaccomplished by localized application. In reference to application ofthe composition, the term “localized”, “local”, or “locally” mean thatthe composition is delivered the targeted area (such as thehypopigmented spot) while minimizing delivery to skin surface notrequiring treatment. The composition may be applied and lightly massagedinto the hypopigmented spot. It is recognized that localized applicationdoes allow for a reasonable amount of the composition to be applied toareas adjacent the hypopigmented spot (i.e., the composition is unlikelyto be applied or to remain within the boundary of the hypopigmented spotwithout some spreading). The form of the composition or thedermatologically acceptable carrier should be selected to facilitatelocalized application. While certain embodiments of the presentinvention contemplate applying a composition locally to a hypopigmentedspot, it will be appreciated that compositions of the present inventioncan be applied more generally or broadly to one or more facial skinsurfaces to reduce the appearance of hypopigmented spots within thosefacial skin regions.

In some embodiments, the composition may be delivered by a variety ofapplicators appropriate for localized and general application. Suchapplicators can include droppers, applicator wands, cotton swabs, or anyother suitable device. Other suitable applicators include SH-0127 penapplicator available from Shya Hsin Plastic Works, Inc., Taiwan andeither the Xpress Tip or liquid filled swab available from SwabPlus,Inc., China. The applicator may be configured to easily apply thecomposition to hypopigmented spots having an approximate diameterbetween about 2 mm and about 10 mm and allowing for a dosed amount ofthe composition of between about 1 to about 50 μL/cm² or between about 1to about 5 μL/cm². In another embodiment, the composition is applied tothe one or more hypopigmented spots and more generally to one or morefacial skin surfaces contemporaneously (i.e., over a period of less than30 minutes or, more typically, less than 5 minutes).

While some methods described herein contemplate applying thecompositions of the present invention with an applicator, it will beappreciated that applicators are not required and the compositions ofthe present invention can also be applied directly by using one's fingeror in other conventional manners.

In one embodiment, the method comprises the steps of applying a firstcomposition comprising an effective amount of a blend of a camelliaserum fraction and a feverfew serum fraction to a hypopigmented spot ora plurality of hypopigmented spots on a skin surface. In anotherembodiment, the method comprises the steps of applying a firstcomposition comprising an effective amount of a blend of a kelp serumfraction and a parsley serum fraction to a hypopigmented spot or aplurality of hypopigmented spots on a skin surface. In anotherembodiment, the method comprises the steps of applying a firstcomposition comprising an effective amount of a combination of a blendof a camellia serum fraction and a feverfew serum fraction, and a blendof a kelp serum fraction and a parsley serum fraction to a hypopigmentedspot or a plurality of hypopigmented spots on a skin surface.Accordingly, the first composition may be any compositions describedherein.

In another embodiment, the method further comprises applying a secondcomposition, which may optionally comprise an effective amount of theserum fraction blends present in the first composition. The first and/orthe second compositions may comprise one or more tone agents, sunscreenactives, anti-inflammatory agents, or optional components. The firstcomposition may be locally applied to the hypopigmented spot orplurality of hypopigmented spots. The second composition may be locallyapplied to the hypopigmented spot or a plurality of hypopigmented spotsto which the first composition is applied or the second composition maybe applied more generally to the skin surface including thehypopigmented spots to which the first composition is applied. Incertain embodiments, the skin surface is facial skin surface whichinclude one or more of the forehead, perioral, chin, periorbital, nose,and cheek skin surfaces. In another embodiment, the first and secondcompositions are applied contemporaneously to at least the cheek,forehead, and chin/perioral skin surfaces. For general application to askin surface and, particularly a facial skin surface, the dosed amountof the first or second composition may be between about 1 to about 50μL/cm² per application (i.e., per single application to the skinsurfaces).

Suitable methods may comprise any one or more of the abovementionedsteps. All of the aforementioned steps are applicable to application,treatment, regulation, and/or improvement of both a single hypopigmentedspot as well as a plurality of hypopigmented spots. Likewise, theexemplary methods that follow are applicable to both a singlehypopigmented spot as well as a plurality of hypopigmented spots.

One suitable method of improving the appearance of a hypopigmented spotincludes the step of topically applying a composition comprising aneffective amount of a blend of a camellia serum fraction and a feverfewserum fraction, and/or a blend of a kelp serum fraction and a parsleyserum fraction to the hypopigmented spot on a skin surface, wherein thecomposition is applied for a period of time sufficient for the serumfractions to improve the appearance of the hypopigmented spot. Anothersuitable method of improving the appearance of hypopigmented spotsincludes the steps of identifying a hypopigmented spot on a skinsurface, applying a composition comprising an effective amount of theserum fraction blend to the hypopigmented spot on the skin surface,wherein the composition is applied for a period of time sufficient forthe serum fractions to improve the appearance of the hypopigmented spot.

Another suitable method is for improving the appearance of apost-inflammatory hypopigmentation. The method may comprise the steps ofidentifying an area of post-inflammatory hypopigmentation on a skinsurface and of applying to the area said skin care compositioncomprising a blend of the serum fractions, which also providesanti-inflammatory effect. An effective amount of said serum fractionsmay be applied at least daily for a period of time sufficient to improvethe appearance of the area of post-inflammatory hypopigmentation. Thecompositions may further comprise a sunscreen active, an additional skintone agent, or combinations thereof.

FORMULATIONS AND EXAMPLES

The following are non-limiting examples of the present invention. Theexamples are given solely for the purpose of illustration and are not tobe construed as limitations of the present invention, as many variationsthereof are possible without departing from the spirit and scope of theinvention, which would be recognized by one of ordinary skill in theart.

In the examples, all concentrations are listed as weight percent, unlessotherwise specified and may exclude minor materials such as diluents,filler, and so forth. The listed formulations, therefore, comprise thelisted components and any minor materials associated with suchcomponents. As is apparent to one of ordinary skill in the art, theselection of these minors will vary depending on the physical andchemical characteristics of the particular ingredients selected to makethe present invention as described herein.

Exemplary Compositions

Table 1 sets forth non-limiting examples of the compositions of thepresent invention. The examples are given solely for the purpose ofillustration and are not to be construed as limitations of the presentinvention, as many variations thereof are possible without departingfrom the spirit and scope of the invention, which would be recognized byone of ordinary skill in the art. In the examples, all concentrationsare listed as weight percent, unless otherwise specified and may excludeminor materials such as diluents, filler, and so forth. The listedformulations, therefore, comprise the listed components and any minormaterials associated with such components. As is apparent to one ofordinary skill in the art, the selection of these minor materials willvary depending on the physical and chemical characteristics of theparticular ingredients selected to make the present invention asdescribed herein.

All examples may be used to treat or improve the appearance of one ormore hypopigmented spots. The present invention may further relate to aregimen involving the localized treatment for one or more hypopigmentedspots by a first composition and/or a more broad or general facial skintreatment by a second composition, which can be applied before or afterthe localized treatment to improve skin tone across the face.

TABLE 1 TABLE 1: Exemplary Compositions Component/% by wt. Ex. A Ex. BEx. C Ex. D Ex. E Ex. F camellia sinensis 1 0.4 1.6 — — — (camelliaserum fraction) (manufactured by IBT*) chrysanthemum 1 1.6 0.4 — — —parthenium (feverfew serum fraction) (manufactured by IBT*) petroselinumcrispum — — — 1 0.4 1.6 (parsley serum fraction) (manufactured by IBT*)macrocystis pyrifera — — — 1 1.6 0.4 (kelp serum fraction) (manufacturedby IBT) N-Acetylglucosamine 0.00 2.00 0.00 0.00 2.00 0.00 Hexamidine0.00 0.09 0.09 0.00 0.09 0.09 Diisethionate Sepiwhite ™ 0.00 0.50 0.500.00 0.50 0.50 (Undecylenoyl- phenylalanine, neutralized) (availablefrom SEPPIC) Sepigel 305 ™ 0.00 2.00 2.00 0.00 2.00 2.00(Polyacrylamide + C13-14 isoparaffin + laureth- 7) (available fromSEPPIC) Dipotassium 0.00 0.10 0.30 0.00 0.10 0.30 GlycyrrhizateHexamidine 0.00 0.09 0.09 0.00 0.09 0.09 Diisethionate Homosalate 0.000.00 9.00 0.00 0.00 9.00 Avobenzone 0.00 0.00 3.00 0.00 0.00 3.00Octocrylene 0.00 0.00 2.60 0.00 0.00 2.60 Oxybenzone 0.00 0.00 1.00 0.000.00 1.00 Octisalate 0.00 0.00 4.50 0.00 0.00 4.50 Butylene Glycol (CAS5.50 5.50 5.50 5.50 5.50 5.50 No. 107-88-0) Niacinamide (CAS No. 5.005.00 5.00 5.00 5.00 5.00 98-92-0) Glycerin (CAS No. 56- 2.50 2.50 2.502.50 2.50 2.50 81-5) DC 1503 Fluid ™ 2.50 2.50 2.50 2.50 2.50 2.50(available from DowCorning) Lubrajel Oil ™(available 1.44 1.44 1.44 1.441.44 1.44 from Sederma) Phenonip XB ™ 1.25 1.25 1.25 1.25 1.25 1.25(available from Clariant) D-panthenol (CAS No. 1.00 1.00 1.00 1.00 1.001.00 81-13-0) Tospearl 2000 ™ 1.00 1.00 1.00 1.00 1.00 1.00(Polymethylsils esquioxane) (CAS No. 68554-70-1) (available from GESilicones/Momentive) DL-Alpha Tocopheryl 0.50 0.50 0.50 0.50 0.50 0.50Acetate (CAS No. 7695- 91-2) Prodew 400 ™ (available 0.50 0.50 0.50 0.500.50 0.50 from Ajinomoto) Pemulen TR-2 ™ 0.25 0.25 0.25 0.25 0.25 0.25(Acrylates/C10-30 Alkyl Acrylate Crosspolymer) (available from Noveon)Polysorbate 20 (CAS No. 0.25 0.25 0.25 0.25 0.25 0.25 9005-64-5) SodiumMetabisulfite 0.25 0.25 0.25 0.25 0.25 0.25 (CAS No. 7681-57-4)Allantoin (CAS No. 97- 0.20 0.20 0.20 0.20 0.20 0.20 59-6) SodiumHydroxide (CAS 0.17 0.17 0.17 0.17 0.17 0.17 No. 1310-73-2) (50%solution by weight in water) Disodium EDTA (CAS 0.10 0.10 0.10 0.10 0.100.10 No. 139-33-3) Xanthan Gum (CAS No. 0.05 0.05 0.05 0.05 0.05 0.0511138-66-2) Sodium Hyaluronate 0.01 0.01 0.01 0.01 0.01 0.01 (CAS No.9067-32-7) Water (CAS No. 7732- QS QS QS QS QS QS 18-5) TOTAL (% byweight of 100.00 100.00 100.00 100.00 100.00 100.00 total composition)*IBT is an abbreviation for Integrated Botanical Technologies, which isnow Akzo Nobel Surface Chemistry LLC, Chicago, Illinois.

According to embodiments of the present invention, the skin carecompositions are generally prepared by conventional methods such as areknown in the art of making topical compositions. Such methods typicallyinvolve mixing of the ingredients in one or more steps to a relativelyuniform state, with or without heating, cooling, application of vacuum,and the like. Typically, emulsions are prepared by first mixing theaqueous phase materials separately from the fatty phase materials andthen combining the two phases as appropriate to yield the desiredcontinuous phase. The compositions are preferably prepared such as tooptimize stability (physical stability, chemical stability,photostability) and/or delivery of the active materials. Thisoptimization may include appropriate pH (e.g., less than 7), exclusionof materials that can complex with the active agent and thus negativelyimpact stability or delivery (e.g., exclusion of contaminating iron),use of approaches to prevent complex formation (e.g., appropriatedispersing agents or dual compartment packaging), use of appropriatephotostability approaches (e.g., incorporation of sunscreen/sunblock,use of opaque packaging), etc.

III. Bio-Activity

Although not wishing to be limited by theory, it is believed thattopical application of various skin tone agents can: (1) stimulate theproduction of melanin in skin melanocytes; and/or (2) interrupt orinhibit an inflammatory cycle. Furthermore, the topical application canlead to evenly toned skin and afford the appearance of younger lookingskin.

One method for predicting the efficacy of the blends of serum fractionsto affect the inflammatory cycle is using in vitro bio-assays. Morespecifically, interleukin signaling inhibition via interleukin-1receptor-associated kinase (IRAK-4) inhibition is the predictivebioassay method described below.

Interleukin-1 Signaling Inhibition: Interleukin-1 (IL-1) is a family ofpro-inflammatory cytokines that initiate biochemical signaling pathwaysto increase inflammation. In the skin, IL-1 is an endogenous factor thatinduces an inflammatory action. It is believed that inhibition of IL-1signaling at any point within its pathways inhibits inflammation.

Interleukin-1 receptor-associated kinase inhibition: Interleukin-1receptor-associated kinase (IRAK-4) is an integral mediator of IL-1signaling that recruits other kinases upon IL-1 stimulation forsubsequent signal transduction. The inhibition of IRAK-4 activity ismeasured by the amount of ATP using the ADP-Glo™ kinase kit (Promega)after incubation of the full-length human recombinant IRAK-4 enzymesystem (Promega) with inhibitors for 30 minutes. The ADP-Glo™ kit wasused in accordance with the manufacturer's instructions.

Melanin synthesis activation: Melanin is the pigment made by melanocytesthat is responsible for skin color. Melanin synthesis only occurs duringthe anagen phase. Prolonging the anagen phase potentially prolongsmelanin synthesis and also provides the opportunity to increase thesynthesis of melanin with specific activators. The activation of melaninis measured in an in vitro model of B16-F1 melanocytes (ATCC) inculture. Melanocytes are incubated with activators for 48 hours and themelanin formed is measured by reading the optical density (O.D.) at 410nm on a spectrophotometer.

Melanin Synthesis Activation Materials: Plates: Corning® 96 Well FlatClear Bottom White Polystyrene TC-Treated Microplates, #3903; Cells:B16-F1 (ATCC); Growing Medium: DMEM, Gibco Invitrogen, #11965-092 with10% FBS and 1% Penicillin/strep/Glutamine (GLBCO cat#15701); serumfractions and blends; preservatives; and controls.

Melanin Synthesis Activation Method: Day 1: Seeding B16-F1 cells,2000/well/100 ul; Day 2: Treating the compounds by adding 10 μl ofdiluted compounds into each well; and Day 4: Measure the color change ineach well. Checking the cell viability under microscope, if cellconfluence is <50%, the data for this point is not used. Measuring ODvalue (melanin product) at V is/UV reader, 410 nM after adding 100 μl of1% NaOH (1 ml 50% NaOH+49 ml H2O) into each well. Note: Due toprevalence of color with serum fractions, wells with media but no cellswere also treated and used as color control blanks to removeover-estimation due to serum color.

Statistical Significance Definition: One-tailed 2-sample t-tests wereconducted. Statistical significance is defined as p<0.05. Examples inthis application are provided with values of p<0.01. Statisticallysignificant and synergistic blends are defined as blends withstatistically significantly greater activity than that of bothindividual serum fraction components.

A. Kelp and Parsley Serum Fractions

(1) The kelp serum fraction, the parsley fraction, and combinations canbe assessed for melanin synthesis action (all serum fractions analyzedat 1% (v/v) of the analytical composition):

TABLE 1 Example 1A: Kelp-Parsley 10:90 Blend K: Lot# MP0726M/MS-0571; P:Lot #PA1117L/HP-0570; K + P: Lot #: B19-0729M/OS-0572 Fold 2-samplet-test vs. blend Serum Fraction (1%) activation (p < 0.01 statisticallysignificant) Kelp 1.62 0.000008 Parsley 1.91 0.000010 Kelp-Parsley 10:906.36 Blend

TABLE 2 Example 1B: Kelp-Parsley 20:80 Blend K: Lot# MP0726M/MS-0571; P:Lot #PA1117L/HP-0570; K + P: Lot #: B19-0729M/OS-0573 Fold 2-samplet-test vs. blend Serum Fraction (1%) activation (p < 0.01 statisticallysignificant) Kelp 1.62 0.000016 Parsley 1.91 0.000022 Kelp-Parsley 20:804.25 Blend

TABLE 3 Example 1C: Kelp-Parsley 30:70 Blend K: Lot# MP0726M/MS-0571; P:Lot #PA1117L/HP-0570; K + P: Lot #: B19-0729M/OS-0574 Fold 2-samplet-test vs. blend Serum Fraction (1%) activation (p < 0.01 statisticallysignificant) Kelp 1.62 0.001324 Parsley 1.91 0.004537 Kelp-Parsley 30:702.66 Blend

TABLE 4 Example 1D: Kelp-Parsley 40:60 Blend K: Lot# MP0726M/MS-0571; P:Lot #PA1117L/HP-0570; K + P: Lot #: B19-0729M/OS-0575 Fold 2-samplet-test vs. blend Serum Fraction (1%) activation (p < 0.01 statisticallysignificant) Kelp 1.62 0.000004 Parsley 1.91 0.000005 Kelp-Parsley 40:604.37 Blend

TABLE 5 Example 1E: Kelp-Parsley 50:50 Blend K: Lot# MP0726M/MS-0571; P:Lot #PA1117L/HP-0570; K + P: Lot #: B19-0729M/OS-0576 Fold 2-samplet-test vs. blend Serum Fraction (1%) activation (p < 0.01 statisticallysignificant) Kelp 1.62 0.000571 Parsley 1.91 0.001749 Kelp-Parsley 50:502.77 Blend

TABLE 6 Example 1F: Kelp-Parsley 70:30 Blend K: Lot# MP0726M/MS-0571; P:Lot #PA1117L/HP-0570; K + P: Lot #: B19-0729M/OS-0578 Fold 2-samplet-test vs. blend Serum Fraction (1%) activation (p < 0.01 statisticallysignificant) Kelp 1.62 0.001198 Parsley 1.91 0.004543 Kelp-Parsley 70:302.61 Blend

TABLE 7 Example 1G: Kelp-Parsley 50:50 Blend K: Lot# 0320J/MS-0363; P:Lot #PA1117L/HP-0499; K + P: Lot #: B19-0215M/OS-0529 Fold 2-samplet-test vs. blend Serum Fraction (1%) activation (p < 0.01 statisticallysignificant) Kelp 2.13 0.000007 Parsley 1.41 0.000007 Kelp-Parsley 50:506.7 Blend

A constant concentration is used to directly demonstrate that the samelevel of the dry matter content of the blended serum fraction hassignificantly more activity than the level of either of the individualserum fractions alone. This demonstrates that the two serums in theblend work synergistically together to afford significantly betterbiological activity. For example in Table 1A, the individual serumfractions of kelp and parsley at 1% give % give 1.62 and 1.91 foldactivation but 1% of a 10:90 blend of these individual serum fractionsgive a significantly and much greater activation of 6.36 fold.

(2) The combination of kelp serum fraction and the parsley serumfraction can be assessed for IRAK-4 inhibition (all serum fractionsanalyzed at 4% (v/v) of the analytical composition):

TABLE 8 Example 1H: Kelp-Parsley 50:50 Blend K: Lot# MP0726M/MS-0571; P:Lot #PA1117L/HP-0570; K + P: Lot #: B19-0729M/OS-0576 % 2-sample t-testvs. blend Serum Fraction (4%) inhibition (p < 0.01 statisticallysignificant) Kelp 67.7 0.001127 Parsley 43 0.000034 Kelp-Parsley 50:5076.3 Blend

TABLE 9 Example 1I: Kelp-Parsley 60:40 Blend K: Lot# MP0726M/MS-0571; P:Lot #PA1117L/HP-0570; K + P: Lot #: B19-0729M/OS-0577 % 2-sample t-testvs. blend Serum Fraction (4%) inhibition (p < 0.01 statisticallysignificant) Kelp 67.7 0.000015 Parsley 43 0.000012 Kelp-Parsley 60:4077.7 Blend

TABLE 10 Example 1J: Kelp-Parsley 70:30 Blend K: Lot# MP0726M/MS-0571;P: Lot #PA1117L/HP-0570; K + P: Lot #: B19-0729M/OS-0578 % 2-samplet-test vs. blend Serum Fraction (4%) inhibition (p < 0.01 statisticallysignificant) Kelp 67.7 0.000336 Parsley 43 0.000022 Kelp-Parsley 70:3076.7 Blend

TABLE 11 Example 1K: Kelp-Parsley 80:20 Blend K: Lot# MP0726M/MS-0571;P: Lot #PA1117L/HP-0570; K + P: Lot #: B19-0729M/OS-0579 % 2-samplet-test vs. blend Serum Fraction (4%) inhibition (p < 0.01 statisticallysignificant) Kelp 67.7 0.001445 Parsley 43 0.000030 Kelp-Parsley 80:2072 Blend

A constant concentration was used to directly demonstrate that the samelevel of dry weight matter in the blended serum fraction hassignificantly more activity than the level of either of the individualserum fractions alone. This shows that the two serums in the blend worksynergistically together to afford significantly better biologicalactivity. For example in Table 11, Experiment 1K, the individual serumfractions of kelp and parsley at 4.0% give 67.7% and 43% inhibitionrespectively, whereas 4.0% of a 50:50 blend of these serum fractionsgives a significantly greater inhibition of 76.3%.

B. Camellia and Feverfew Serum Fractions

(1) The camellia serum fraction, the feverfew fraction, and combinationscan be assessed for melanin synthesis action (all serum fractionsanalyzed at 0.8% (v/v) of the analytical composition):

TABLE 12 Example 2A: Camellia-Feverfew 50:50 Blend C:Lot#TECJ062904-0161-01; F: Lot#FF0811K/TL; C + F: Lot#B02-0712M/OS-0561Fold 2-sample t-test vs. blend Serum Fraction (1%) activation (p < 0.01statistically significant) Camellia 2.26 0.003412 Feverfew 1.72 0.000648Camellia-Feverfew 50:50 3.24 Blend

TABLE 13 Example 2B: Camellia-Feverfew 80:20 Blend C:Lot#TECJ062904-0161-01; F: Lot#FF0811K/TL; C + F: Lot#B02-0719M/OS-0567Fold 2-sample t-test vs. blend Serum Fraction (1%) activation (p < 0.01statistically significant) Camellia 2.26 0.000309 Feverfew 1.72 0.000088Camellia-Feverfew 80:20 3.73 Blend

TABLE 14 Example 2C: Camellia-Feverfew 20:80 Blend C:Lot#TECJ062904-0184; F: Lot#FF0811K/TL; C + F: Lot#FFCSWASC02P-0351 Fold2-sample t-test vs. blend Serum Fraction (1%) activation (p < 0.01statistically significant) Camellia 2.26 0.000041 Feverfew 2.02 0.000003Camellia-Feverfew 20:80 5.02 Blend

A constant concentration is used to directly demonstrate that the samelevel of the dry weight matter in the blended serum fraction hassignificantly more activity than the level of either of the individualserum fractions alone. This demonstrates that the two dry weight mattersin the blend work synergistically together to afford significantlybetter biological activity. For example in Table 14, Experiment 2C, theindividual serum fractions of camellia and feverfew at 1% give 2.26 and2.02 fold activation respectively, whereas 1% of a 20:80 blend of theseserum fractions give a significantly and much greater activation of 5.02fold. The same is demonstrated for a 50:50 blend (Table 12, Experiment2A) and 80:20 blend (Table 13, Experiment 2B).

(2) The camellia serum fraction, the feverfew fraction, and combinationscan be assessed for IRAK-4 inhibition (all serum fractions analyzed at0.8% (v/v) of the analytical composition):

TABLE 15 Example 2D: Camellia-Feverfew 50:50 Blend C:Lot#TECJ062904-0161-01; F: Lot#FF0811K/TL; C + F: Lot#B02-0712M/OS-0561% 2-sample t-test vs. blend Serum Fraction (0.8%) inhibition (p < 0.01statistically significant) Camellia 8 0.000604 Feverfew 9 0.000502Camellia-Feverfew 50:50 37.7 Blend

TABLE 16 Example 2E: Camellia-Feverfew 80:20 Blend C:Lot#TECJ062904-0161-01; F: Lot#FF0811K/TL; C + F: Lot#B02-0719M/OS-0567% 2-sample t-test vs. blend Serum Fraction (0.8%) inhibition (p < 0.01statistically significant) Camellia 8 0.000045 Feverfew 9 0.000029Camellia-Feverfew 80:20 54.3 Blend

A constant concentration is used to directly demonstrate that the samelevel of the dry weight matter in the blended serum fraction hassignificantly more activity than the level of either of the individualserum fractions alone. This demonstrates that the two serums in theblend work synergistically together to afford significantly betterbiological activity. For example in Table 16, Example 2E, the individualserum fractions of camellia and feverfew at 0.8% give 8% and 9%inhibition respectively, whereas 0.8% of a 80:20 blend of these serumfractions gives a significantly and much greater inhibition of 54.3%.

The dimensions and values disclosed herein are not to be understood asbeing strictly limited to the exact numerical values recited. Instead,unless otherwise specified, each such dimension is intended to mean boththe recited value and a functionally equivalent range surrounding thatvalue. For example, a dimension disclosed as “40 mm” is intended to mean“about 40 mm.”

All documents cited in the Detailed Description of Embodiments of theInvention are, in relevant part, incorporated herein by reference; thecitation of any document is not to be construed as an admission that itis prior art with respect to the present invention. To the extent thatany meaning or definition of a term in this document conflicts with anymeaning or definition of the same term in a document incorporated byreference, the meaning or definition assigned to that term in thisdocument shall govern.

While particular embodiments of the present invention have beenillustrated and described, it would be obvious to those skilled in theart that various other changes and modifications can be made withoutdeparting from the spirit and scope of the invention. It is thereforeintended to cover in the appended claims all such changes andmodifications that are within the scope of this invention.

What is claimed is:
 1. A bioactive composition having anti-agingactivity, said bioactive composition comprising: a. an effective amountof: i. a first composition comprising a combination of a camellia serumfraction and a feverfew serum fraction in a weight ratio of 10:90 toabout 90:10; and/or ii. a second composition comprising a combination ofa parsley serum fraction and a kelp serum fraction in a weight ratio of10:90 to about 90:10.
 2. The bioactive composition of claim 1, whereinthe first composition comprises: a. from about 0.001 wt % to about 15 wt% of the camellia serum fraction; and b. from about 0.001 wt % to about15 wt % of the feverfew serum fraction, wherein the wt % is based on thetotal weight of the composition.
 3. The bioactive composition of claim2, wherein a weight ratio of the camellia serum fraction to the feverfewserum fraction ranges from about 20:80 to about 80:20.
 4. The bioactivecomposition of claim 2, wherein a weight ratio of the camellia serumfraction to the feverfew serum fraction is about 50:50 to about 80:20.5. The bioactive composition of claim 2, further comprising a thickeningagent.
 6. The bioactive composition of claim 1, wherein a weight ratioof the parsley serum fraction to the kelp serum fraction ranges fromabout 30:70 to about 70:30.
 7. The bioactive composition of claim 1,wherein a weight ratio of the parsley serum fraction to the kelp serumfraction is about 50:50.
 8. The bioactive composition of claim 1,further comprising a thickening agent.
 9. The bioactive composition ofclaim 1 comprising a combination of the first composition and the secondcomposition.
 10. The bioactive composition of claim 9, wherein the firstcomposition comprises: a. from about 0.001 wt % to about 15 wt % of thecamellia serum fraction, and b. from about 0.001 wt % to about 15 wt %of the feverfew serum fraction; and wherein the second compositioncomprises: c. from about 0.001 wt % to about 15 wt % of the parselyserum fraction, and d. from about 0.001 wt % to about 15 wt % of thekelp serum fraction, wherein the wt % is based on the total weight ofthe composition.
 11. The bioactive composition of claim 1, wherein saidbioactive composition further possesses one or more of the followingproperties: antioxidant and/or free radical scavenging, inhibition ofenzymes associated with skin aging and environmental damage, skinprotection, repair of human tissue or skin, UV protection, interruptionof an inflammatory cycle, darkening regions of mammalian skin, reducingthe appearance of skin hypopigmentation, stimulating one or more stepsin melanin synthesis, protection from ozone and/or protection fromosmotic shock.
 12. The bioactive composition of claim 1, wherein saidanti-aging activity comprises hair and/or skin anti-aging activity. 13.The bioactive composition of claim 11 wherein said bioactive compositioninhibits enzymes associated with skin aging and environmental damage.14. The bioactive composition of claim 13 wherein said bioactivecomposition inhibits melanogenesis enzyme activity.
 15. The bioactivecomposition of claim 13 wherein said bioactive composition inhibitstrypsin activity, tyrosinase activity, or both.
 16. The bioactivecomposition of claim 11, wherein said composition increases anti-oxidantactivity, free radical scavenging activity, or both.
 17. The bioactivecomposition of claim 1 wherein said bioactive composition interrupts aninflammatory cycle.
 18. The bioactive composition of claim 11 whereinsaid bioactive composition is effective in darkening regions ofmammalian skin.
 19. The bioactive composition of claim 11 wherein saidbioactive composition is effective in reducing the appearance of skinhypopigmentation.
 20. The bioactive composition of claim 11 wherein saidbioactive composition is effective in stimulate one or more steps inmelanin synthesis.